LOCUS       sequence-289829-        8400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of CeAIN2_254-400-DmGW182_1-634-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8400)
  AUTHORS   Huntzinger E, Kuzuoglu-Ozturk D, Braun JE, Eulalio A, Wohlbold L, 
            Izaurralde E
  TITLE     The interactions of GW182 proteins with PABP and deadenylases are 
            required for both translational repression and degradation of miRNA 
            targets.
  JOURNAL   Nucleic Acids Res. 2013 Jan;41(2):978-94. doi: 10.1093/nar/gks1078. 
            Epub 2012 Nov 21.
  PUBMED    23172285
REFERENCE   2  (bases 1 to 8400)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1093/nar/gks1078"; journalName: "Nucleic Acids Res"; date: 
            "2013-01"; volume: "41"; issue: "2"; pages: "978-94"
COMMENT     Sequence Label: pAc5.1B-EGFP-CeAIN2_254-400-DmGW182_1-634-dsRN...
FEATURES             Location/Qualifiers
     source          1..8400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2633..2654)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2894..2913)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3241..3262
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5685..5726
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5736..5753
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5776..5793)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5826..5960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5850..5869)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5904..5923
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6582..7170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6662..6681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7341..8132,8133..8201))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7964..7983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8202..8306)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8374..8392
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
      301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
      361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
      421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
      481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
      541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
      601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
      661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
      721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
      781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
     1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
     1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
     1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
     1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
     1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
     1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
     1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
     1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
     1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
     1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
     1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
     2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg
     2581 taccaacatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2641 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2701 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2761 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2821 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2881 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2941 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3001 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3061 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3121 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3181 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3241 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3301 caagccaagc ttaatggatg attattcaat gtggactgat gagaatgatg aagacgcgaa
     3361 aaagaaaaaa tcgttgagag ataagggatt gagtggatgg ggtgatgctg atgtatcaaa
     3421 cagtaaacca attcgccgtt ggattgtgcc agaaggccaa gaagaagact ttgaaacggc
     3481 aatggaacga tgtccaagtc atttgaaaaa gaaagcattg aatgaagaag ctcttcgtcg
     3541 acgtttaaca tctgaaaatc cacaaatagt ggcacaagct caacaacaag ccgaagaaga
     3601 ggcgaatgct atgatgagga ttggacgaag acccattgtt cccagtggat ggggtgatat
     3661 tccatctgaa attagtactg acaagtcaga attcgatcaa agctcgtcac gtggttggga
     3721 tgaaggaagt ataagcggtg gtggaagttg cggccgcatg cgtgaagccc ttttttccca
     3781 agatggctgg ggctgtcagc atgttaacca ggatactaat tgggaagttc ccagttcgcc
     3841 agaaccagcc aataaggatg cacccggtcc accaatgtgg aagccaagca ttaacaatgg
     3901 tactgatctt tgggagtcca atttgagaaa cggaggtcag ccggccgcac agcaagttcc
     3961 aaagccgtcg tggggtcata caccatcctc taacttaggt ggaacatggg gtgaggacga
     4021 cgatggcgcc gatagtagta gtgtgtggac tggaggagct gttagcaacg cgggatccgg
     4081 agctgcagtg ggagtaaacc aagccggagt taatgtcggt ccaggcggtg ttgtttcgtc
     4141 tggcggacct cagtggggac aaggtgtcgt tggcgtcgga cttggatcaa ctggcggaaa
     4201 tggttccagt aacattacag gctcttcggg cgttgccacg ggaagcagtg gcaattcaag
     4261 taatgccgga aatggatggg gcgatccacg cgagattcgt ccactgggcg tcggaggatc
     4321 gatggacatc cgtaacgtcg agcaccgtgg aggaaatgga tcaggcgcca catccagtga
     4381 cccgcgtgat atccgtatga ttgacccacg cgatccaatc cggggcgacc cacgcggcat
     4441 ttcaggacgc ctcaacggta cgtcagagat gtggggacac cacccgcaga tgtcgcacaa
     4501 tcaactgcag ggaattaata agatggtcgg acagagcgtt gcgacagcta gtacgagcgt
     4561 tggcacgtca ggatccggaa ttggaccagg cggaccggga ccaagcacgg tttcgggaaa
     4621 cattccgacg caatggggtc cagcccagcc cgttagtgtc ggagttagcg gaccgaagga
     4681 tatgtcgaag caaattagcg gctgggaaga gccgtcgcct cccccacaac gccgtagcat
     4741 cccaaattat gacgacggaa cgtccctctg gggacaacag acacgcgtcc cggccgctag
     4801 tggacattgg aaggatatga cagactccat tggacgcagc agccacctga tgcgcggaca
     4861 gagtcagacc ggcggaattg gcattgctgg agttggaaat agcaatgttc cagtgggagc
     4921 caatccaagt aatcctataa gcagtgtagt tggacctcaa gcccggattc catctgtggg
     4981 cggcgtacaa cacaaaccag acggcggcgc tatgtgggtg cattccggca atgtaggtgg
     5041 cagaaataat gttgctgctg ttactacttg gggagatgac actcatagcg ttaatgtcgg
     5101 cgctcccagc agtggcagtg tatccagcaa caattgggtt gatgacaagt ccaactcaac
     5161 cttggcacaa aactcttgga gcgacccggc ccctgttgga gttagttggg gcaataagca
     5221 aagcaaaccg ccaagcaata gtgcttcatc aggttggagc actgctgcgg gcgtggtgga
     5281 tggggttgat ctaggatctg agtggaacac gcacgggggg attattggaa aatctcagca
     5341 acaacaaaaa ctagcgggac ttaacgtggg aatggtgaac gtaattaacg cggagatcat
     5401 taagcaaagc aagcaataca ggatccttgt cgagaacggc tttaaaaagg aagatgtaga
     5461 gcgggcatta gtgattgcta atatgaacat cgaagaggca gccgatatgc tccgtgccaa
     5521 ctcatcccta tcaatggatg gttggcgtcg acatgatgag tcccttggat cttatgccga
     5581 ccacaatagt tcaacaagca gcggtggatt tgctggtcgt tacccggtca acagtggaca
     5641 accttcaatg tcctttcctt aatctagagg gcccgcggtt cgaaggtaag cctatcccta
     5701 accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt
     5761 aaacccgctg atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga
     5821 tcgaggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5881 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     5941 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     6001 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
     6061 atgatcagtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     6121 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
     6181 caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg
     6241 atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc
     6301 actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc
     6361 gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt
     6421 atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg
     6481 caggtagatg acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccag
     6541 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6601 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6661 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6721 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6781 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6841 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6901 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6961 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7021 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7081 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7141 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7201 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7261 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7321 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7381 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7441 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7501 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7561 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7621 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     7681 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7741 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7801 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7861 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7921 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     7981 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8041 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8101 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8161 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8221 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8281 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     8341 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
//
